影响枯草芽胞杆菌和荧光假单胞菌原生质体再生的因素

目的：为了提高再生率,对影响革兰阳性菌枯草芽胞杆菌KR株和革兰阴性菌荧光假单胞菌B13株原生质体再生的因素进行研究。方法：研究了酶解时间,再生方式,再生培养基中稳定剂的种类,Ca^2＋、Mg^2＋、琥珀酸钠、L-色氨酸的浓度及培养基的放置时间对KR和B13株原生质体再生的影响。结果：对KR株酶解20min,采用夹层培养,再生培养基中加入0.6mol/L蔗糖、0.03mol/L Ca^2＋、0.02mol/L Mg^2＋、0.3mol/L琥珀酸钠、0.2mol/L L-色氨酸,培养基在37℃放置72h,原生质体再生率可达42.7%;对B13酶解15min,采用夹层培养,培养基中加入0.6mol/L NaCl、0.02mol/L Ca^2＋、0.01mol/L Mg^2＋、0.3mol/L琥珀酸钠、0.1mol/L L-色氨酸,培养基在37℃放置48h,原生质体再生率可达15.3%。结论：影响革兰阳性菌枯草芽胞杆菌KR株和革兰阴性菌荧光假单胞菌B13株原生质体再生的因素是不同的。

Influencing Factors of Protoplast Regeneration of Bacillus subtilis and Pseudomonas fluorescent

Objective： In order to improve the regeneration rate, the effects of different factors on the regeneration of Gram-positive Bacillus subtilis KR strain and Gram-negative Pseudomonas fluorescent B13 strain were conducted. Meth. otis： The experiments were designed for pretoplast regeneration of KR and B13 strains as the following factors： lysozyme treatment time, regeneration mode, stabler in culture medium, and the concentration of Ca^2＋, Mg^2＋, sodium succinate acid, L-tryptophan, and culture medium placed time. Results： The optimal protoplast regeneration conditions for KR strain were lysozyme treatment 20 min, interlayer, 0.6 mol/L sucrose, 0.03 mol/L Ca^2＋, 0.02 mol/L Mg^2＋, 0.3 mol/L sodium succinate acid, 0.2 mol/L L-tryptophan and culture medium placed 72 h under 37℃. The optimal protoplast regeneration conditions for B13 strain were lysozyme treatment 15 min, interlayer, 0.6 mol/L NaC1, 0.02 mol/L Ca^2＋, 0.01 mol/L Mg^2＋, 0.3 mol/L sodium succinate acid, 0.1 mol/L L-tryptophan and culture medium placed 48 h under 37℃. The highest protoplast regeneration rates of KR strain and B13 strain were 42.7% and 15.3% respectively. Conclusion： The factors which affected Gram-positive bacterium＇s protoplast regeneration were different from which affected Gram-negative bacterium.